胚胎干细胞(ESC)衍生物为构建组织工程皮肤等效物(TESE)提供了巨大的希望。然而,ESC的收获会破坏可行的胚胎,并可能导致对其应用的政治和伦理关注。在目前的研究中,我们指导小鼠孤雌生殖胚胎干细胞(pESCs)分化为成纤维细胞,建造的TESE,并评价其在体内的功能。
测试了pESC的干性标记表达和多能分化能力。胚状体(EB)形成和贴壁培养后,富集间充质干细胞(MSC)并定向分化为成纤维细胞谱系。通过定量实时PCR和ELISA评估衍生的成纤维细胞的特征。通过小鼠皮肤缺损修复模型测试构建的TESE的功能能力。
小鼠pESCs表达干性标记,可形成含有三个胚层的畸胎瘤。MSC可以从EBs的外植体中富集并定向分化成成纤维细胞谱系。这些细胞表达高水平的生长因子,包括FGF,EGF,VEGF,TGF,PDGF,和IGF1,类似于ESC来源的成纤维细胞和小鼠成纤维细胞。接种到胶原蛋白凝胶中,pESCs来源的成纤维细胞可以形成TESE。由pESCs来源的成纤维细胞构建的TESE移植后15天可以成功修复小鼠皮肤缺损。
pESCs可以诱导分化为成纤维细胞谱系,可应用于TESE的构建和皮肤缺损的修复。特别是,pESC衍生品避免了政治和道德问题的限制,并为再生医学提供了有希望的来源。
Embryonic stem cell (ESC) derivatives hold great promise for the construction of tissue-engineered skin equivalents (TESE). However, harvesting of ESCs destroys viable embryos and may lead to political and ethical concerns over their application. In the current
study, we directed mouse parthenogenetic embryonic stem cells (pESCs) to differentiate into fibroblasts, constructed TESE, and evaluated its function in vivo.
The stemness marker expression and the pluripotent differentiation ability of pESCs were tested. After embryoid body (EB) formation and adherence culture, mesenchymal stem cells (MSCs) were enriched and directed to differentiate into fibroblastic lineage. Characteristics of derived fibroblasts were assessed by quantitative real-time PCR and ELISA. Functional ability of the constructed TESE was tested by a mouse skin defects repair model.
Mouse pESCs expressed stemness marker and could form teratoma containing three germ layers. MSCs could be enriched from outgrowths of EBs and directed to differentiate into fibroblastic lineage. These cells express a high level of growth factors including FGF, EGF, VEGF, TGF, PDGF, and IGF1, similar to those of ESC-derived fibroblasts and mouse fibroblasts. Seeded into collagen gels, the fibroblasts derived from pESCs could form TESE. Mouse skin defects could be successfully repaired 15 days after transplantation of TESE constructed by fibroblasts derived from pESCs.
pESCs could be induced to differentiate into fibroblastic lineage, which could be applied to the construction of TESE and skin defect repair. Particularly, pESC derivatives avoid the limitations of political and ethical concerns, and provide a promising source for regenerative medicine.